A reactivity-based probe of the intracellular labile ferrous iron pool

Benjamin Spangler, Charles W. Morgan, Shaun D. Fontaine, Mark N.Vander Wal, Christopher J. Chang, James A. Wells, Adam R. Renslo

Research output: Contribution to journalArticlepeer-review

123 Scopus citations

Abstract

Improved methods for studying intracellular reactive Fe(II) are of significant interest for studies of iron metabolism and disease-relevant changes in iron homeostasis. Here we describe a highly selective reactivity-based probe in which a Fenton-Type reaction with intracellular labile Fe(II) leads to unmasking of the aminonucleoside puromycin. Puromycin leaves a permanent and dose-dependent mark on treated cells that can be detected with high sensitivity and precision using a high-content, plate-based immunofluorescence assay. Using this new probe and screening approach, we detected alteration of cellular labile Fe(II) in response extracellular iron conditioning, overexpression of iron storage and/or export proteins, and post-Translational regulation of iron export. We also used this new tool to demonstrate that labile Fe(II) pools are larger in cancer cells than in nontumorigenic cells.

Original languageEnglish (US)
Pages (from-to)680-685
Number of pages6
JournalNature Chemical Biology
Volume12
Issue number9
DOIs
StatePublished - Sep 1 2016
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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